1. Field of the Invention
The present invention is related to a cultivation of strains of Bacillus polyfermenticus. More particularly, the present invention is related to a large-scale production of Bacillus polyfermenticus strains in an economic way with a high yield.
2. Description of the Prior Arts
Beneficial bacteria, which have been used for medical purpose and for medicines for treating intestinal disorders, include strains of Bifidobacterium, Lactobacillus, Enterococcus, Clostridium butyricum, Lactobacillus sporogenes, Bacillus subtilis, Bacillus polyfermenticus and the like. In particular, B. Polyfermenticus which is commonly called as "Bispan" strains, have been appropriately used for the treatment of long-term intestinal disorders, since the live strains in the form of active endospores can successfully reach the target intestine.
Bispan strains are stated in Japanese Pharmacopoeia as a amylolytic bacillus along with Bacillus subtilis and Bacillus mesentericus. See Japanese Pharmacopoeia, Medicine Composition Standard Standard 1005-1007 (1993). However, the term "Bispan" does not appear in the International Nomenclature such as Bergey's Manual of Systematic Bacteriology (Sneath, P.H.A.: Endospore-forming gram-positive rods and cocci. In Bergey's Manual of Systematic Bacteriology 1st ed. 1104-1139, Williams & Wilkins, Baltimore (1986)). Several studies suggested that the Bispan strains are significantly similar to Bacillus subtilis strains in terms of morphological and biochemical properties. However, the Bispan strains are still distinct from Bacillus subtilis strains that the former is capable of metabolizing lactose and produces a larger amount of acetic acid and lactic acid from glucose and lactose, respectively, than the latter does.
The Bispan strain, a endospore-forming rod, was for the first time isolated from air by Dr. Terakado in 1933. It produces a variety of enzymes which lysis pathogenic strains such as typhoid bacillus, paratyphoid bacillus, shigella, cholera. The uptake of the Bispan strains can enhance the appetite and promote digestion action in the human by serving as vitamin B.sub.1 and B.sub.2 sources as well as strengthens the protection from non-oral infection and oral immunization.
The cultivation and application of the Bispan strain was described in Japanese patent publication No. sho 11-123 "A process for manufacturing medicine for intestine disorders by using bacteria." It teaches that the medicine for intestine disorders are manufactured by carrying out four (4) cycles of procedure consisted of heating a Bispan culture after the cultivation to 65.degree. C. for 1 hour, allowing to stand the heated culture at 33.degree. C. for 24 hours and then heating it to 65.degree. C. for 1 hour; making the culture weak acidic using acetic acid and dehydrating it under an elevated temperature; adding 0.3-0.5% azueous sulfuric acid and allowing the mixture to stand at 37.degree. C.; dehydrating the mixture under an elevated temperature; and adding a certain amount of starch and lactose to the thus-treated mixture to produce the medicine for intestinal disorders.
The known cultivation of Bispan strain has a shortcoming that it requires too many steps to harvest the cells from the culture, resulting in an increase of the production cost. Further, it uses a very expensive culture medium such as Bouillon medium.
Therefore, it had been needed a large-scale process for cultivating the Bispan strain and harvesting its endospores with a high yield in an economic way.